NEW SUMOylation 1 Affinity Beads

SUMO1 Antibody Mouse Monoclonal (30 assays) (Agarose bead conjugate)
$0.00
SKU
ASM11-beads

As part of the Signal-Seeker™ product line, SUMOylation 1 affinity beads have been optimized in order to detect endogenous levels of SUMO1 modified proteins, which often represent <1% of the target protein. SUMOylation 1 affinity beads comprise an anti-SUMO1 antibody (Clone 7A1A2) that has been chemically conjugated to Protein G beads. Validation studies done in HAP1 wildtype and SUMO1 knockout cells have shown that these beads can immunoprecipitate a wide range of SUMO 2/3 modified proteins without detectable leaching of either heavy or light chains in an IP assay. A comprehensive Signal-Seeker™ SUMOylation 2/3 Detection Kit is also available (BK162) and is recommended for first time users.

Validation Data: see datasheet

Each lot of affinity-bead is quality controlled  to provide high batch to batch consistency, see COA documents.


Immunoprecipitating total SUMO1 profiles using the Signal-Seeker™ SUMOylation 1 Affinity Beads  compared to older SUMO1 tools

(A) HAP1 wildtype (WT) or SUMO1 knockout (KO) lysate, was obtained using BlastR lysis and filter system. 1 mg of each lysate were incubated with 40 µg of each SUMO1 affinity reagent: ASM11-beads (Cytoskeleton), 21C7 (Invitrogen—purified), 21C7 (DSHB—supernatant), D11-beads (Santa Cruz) and conjugated SUMO1 IgG control beads (CIG03).  21C7 antibodies were captured with protein G agarose beads to enrich for SUMO-1 modified proteins. Samples were separated by SDS-PAGE and transferred to PVDF.  Enriched SUMO1 samples were analyzed by western blot using ASM01 (Cytoskeleton) antibody at 1:5000, and mouse Trubelot Ultra-HRP secondary at 1:1000 in 5% milk. Trueblot secondary was used to minimize heavy and light chain detection from 21C7 samples. 

(B): IP was performed using ASM11 as in Fig 1A.  SUMO1 modified proteins were visualized with ASM01 1:5000, and anti-mouse secondary at 1:20,000 to highlight the profile of SUMOylated proteins in the region between 64-30 kDa that may be masked by heavy and light chain interference when using unconjugated antibodies for IP.

Immunoprecipitating SUMO1 modified target proteins using the Signal-Seeker™ SUMOylation 1 Affinity Beads  compared to older SUMO1 tools

HAP1 wildtype (WT) or SUMO1 knockout (KO) lysate, was obtained using BlastR lysis and filter system. 1 mg of each lysate were incubated with 40 µg of each SUMO1 affinity reagent: ASM11-beads (Cytoskeleton), 21C7 (DSHB—supernatant), and conjugated SUMO1 IgG control beads (CIG03).  21C7 antibodies were captured with protein G agarose beads to enrich for SUMO-1 modified proteins. Samples were separated by SDS-PAGE and transferred to PVDF.  Target proteins: (A) TFII-I, RanGAP1, and (B) schmd1 were analyzed for their SUMO1 modified forms by western blot. Anti-rabbit-HRP labeled secondary antibody was used at 1:10,000. All three primary antibodies are rabbit polyclonal antibodies, and should not bind heavy and light chain fragments from the IP antibody.

Amount:
Each package contains enough SUMOylation 1 beads for 30 reactions.

 

For more information contact:  signalseeker@cytoskeleton.com

Associated Products:

Signal-Seeker™ SUMOylation 1 Detection Kit (Cat. # BK165)

Signal-Seeker™ SUMOylation 2/3 Detection Kit (Cat. # BK162)

Signal-Seeker™ Ubiquitination Detection Kit (Cat. # BK161)

Signal-Seeker™ Phosphotyrosine Detection Kit (Cat. # BK160)

Signal-Seeker™: BlastR™ Rapid Lysate Prep Kit (Cat. # BLR01)

Signal-Seeker™: PTMtrue™ SUMOylation 1 Antibody (Cat.# ASM01)

For product Datasheets and MSDSs please click on the PDF links below.

Click on the pdf icon below to download the datasheet

Certificate of Analysis:  Lot 021

New product released in 2018!   For the most recent publications citing this and other Signal-Seeker™ products, see our Signal-Seeker™ Validation Data Page click here

Visit our Signal-Seeker™ Tech Tips and FAQs page for technical tips and frequently asked questions regarding this and other Signal-Seeker™ products click here

 

If you have any questions concerning this product, please contact our Technical Service department at tservice@cytoskeleton.com